Hi,
I would like to look at two different proteins with broad MW (10 and 100KDa) on the same gel. I looked on YouTube and found a video below where you can pour the high and low gradient SDS gels in the same cast. I am just wondering if anyone uses this technique to share their experience and see how the bands look like before I risk my precious samples, please?
The video
https://www.youtube.com/watch?v=zu5a-kpMK8k
Any response appreciated
I prefer to use gradient gels over isogradic gels and have run literally hundreds of them - all large format (i.e., 16 x 16 cm gels). You may be able to buy, rather than experiment with pouring, gradient gels in the format that fits your electrophoresis cell. Before you commit your sample, I would suggest you just run standards and, perhaps, BSA (67 kDa) to see if you are getting the results you want. I can provide you with a recipe for these gels, but you will also need a gradient former - most labs don't have these lying around, and they are quite expensive (>$200) t0 buy.
Many Thanks Prof Allen for your reply.
It would be appreciated please if I can have ur protocol so I will be more confident performing the experiment.
Kind regards
Hi, I have experienced high and low gradient SDS gels in the same cast, only for small gel, as in the video. It is not difficult, you have only to do some test before your experiments. As said belove in general it is better to buy precast gel but, if you can not buy them, I assure you that the handmade gels give you good results.
If you need some tips you can send me a message anytime.
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